(296 days)
The Omega Laboratories Hair Drug Screening Assay (Opiates, Oxycodone and Hydrocodone) is an in vitro diagnostic test that is intended for the qualitative detection of opiates (calibrated with morphine) and oxycodone and hydrocodone (calibrated with oxycodone) at or above 300 pg/mg in human head and body hair. To confirm a screen positive result. a more specific alternate chemical method, such as Gas ChromatographyMass Spectrometry (GC/MS) operating in the selected ion monitoring (SIM) mode is the preferred method with deuterated internal standards. Professional judgment should be applied to any drug of abuse test result, particularly when presumptive positive results are obtained. This test is intended exclusively for single laboratory use only and is not intended for sale to anyone.
The Omega Laboratories Hair Drug Screening Assays are test systems that utilize ELISA assays for the qualitative detection of morphine and related opiates (calibrated with morphine) and oxycodone and hydrocodone (calibrated with oxycodone) at or above 300 pg/mg in head hair samples. The Omega Laboratories Hair Drug Screening Assay for Opiates, Oxycodone and Hydrocodone provide only preliminary analytical test results. A more specific alternate chemical method must be used in order to obtain a confirmed result. Gas Chromatograph - Mass Spectrometry operating in the selected ion monitoring (SIM) mode or GC/MS/MS in selected reaction mode (SRM) is the preferred method with deuterated internal standards.
Here's a summary of the acceptance criteria and the study that proves the device meets them, based on the provided text:
Device Name: Omega Laboratories Hair Drug Screening Assay (Opiates, Oxycodone and Hydrocodone)
Indications for Use: The device is intended for the qualitative detection of opiates (calibrated with morphine) and oxycodone and hydrocodone (calibrated with oxycodone) at or above 300 pg/mg in human head and body hair. It's for single laboratory use only and not for sale to anyone. A more specific alternate chemical method, such as Gas Chromatography/Mass Spectrometry (GC/MS) operating in the selected ion monitoring (SIM) mode, is the preferred method for confirmation of screen positive results.
1. Table of Acceptance Criteria and Reported Device Performance
The document doesn't explicitly state "acceptance criteria" as a set of quantified thresholds. Instead, it demonstrates performance studies to show "substantial agreement" with a predicate device and ensures reliable qualitative detection at the cutoff. Based on the provided performance studies, here's an interpretation of the implied acceptance and the reported performance.
Implicit Acceptance Criterion: The device should consistently and accurately identify positive and negative samples around the cutoff concentration (300 pg/mg) and show substantial agreement with GC/MS confirmation. It should also demonstrate precision, specificity, and stability under various conditions.
| Performance Characteristic | Implied Acceptance Criteria (Interpretation) | Reported Device Performance (Summary) |
|---|---|---|
| Precision (Intra-assay) | All samples below cutoff should be negative; all samples above cutoff should be positive. Samples at the cutoff and slightly below/above may show variability, but consistency is key. | For Opiates and Oxycodone, all samples from 0 to 225 pg/mg were negative, and all from 375 to 600 pg/mg were positive (11/11). For Hydrocodone, 0-225 pg/mg were negative (except one sample at 225 pg/mg was positive in 1 replicate), and 375-600 pg/mg were positive (10/10). |
| Precision (Inter-assay) | High consistency (all negative below cutoff, all positive above cutoff) over multiple days. | For Opiates and Oxycodone, all samples from 0 to 225 pg/mg were negative (220/220), and all from 375 to 600 pg/mg were positive (220/220). For Hydrocodone, 0 to 150 pg/mg were negative (100/100), and 375 to 600 pg/mg were positive (100/100). At 225 pg/mg (75% below cutoff), 61% were negative and 39% positive, demonstrating variability near cutoff. |
| Agreement with GC/MS (Opiates) | High concordance with GC/MS, especially for high positive and negative samples. Few discordant results, particularly near cutoff. | Out of 226 samples: - 208/208 (100%) agreement for Negative (70), < 50% cutoff (4), and > 50% above cutoff (116) by GC/MS. - Near Cutoff Negative (9): 2 Positive by ELISA, 7 Negative by ELISA. - Near Cutoff Positive (24): 24 Positive by ELISA, 1 Negative by ELISA. Overall strong agreement, with expected variability near cutoff. Discordant samples were mostly near the cutoff. |
| Agreement with GC/MS (Oxycodone) | High concordance with GC/MS, especially for high positive and negative samples. Few discordant results, particularly near cutoff. | Out of 483 samples (after exclusions): - 361/361 (100%) agreement for Negative (140), < 50% cutoff (6), and > 50% above cutoff (221) by GC/MS. - Near Cutoff Negative (14): 6 Positive by ELISA, 8 Negative by ELISA. - Near Cutoff Positive (94): 94 Positive by ELISA, 2 Negative by ELISA. Overall strong agreement, with expected variability near cutoff. Discordant samples were mostly near the cutoff. |
| Agreement with GC/MS (Hydrocodone) | High concordance with GC/MS, especially for high positive and negative samples. Few discordant results, particularly near cutoff. | Out of 500 samples (after exclusions): - 343/343 (100%) agreement for Negative (142), < 50% cutoff (8), and > 50% above cutoff (201) by GC/MS. - Near Cutoff Negative (25): 8 Positive by ELISA, 17 Negative by ELISA. - Near Cutoff Positive (110): 110 Positive by ELISA, 6 Negative by ELISA. Overall strong agreement, with expected variability near cutoff. Discordant samples were mostly near the cutoff. |
| Cross-Reactivity | Minimal cross-reactivity with unrelated compounds. Structurally similar compounds may show some cross-reactivity as expected, but this should be characterized. | Detailed tables provided showing percent cross-reactivity for numerous compounds. Structurally similar opiates/opioids show varying degrees of cross-reactivity. Unrelated compounds generally showed no interference at high concentrations (e.g., NEG result at -50% CO for many compounds, turning POS at +125% and +150% CO, indicating no false positive at low concentrations). Specific opiate/oxycodone related compounds did show cross-reactivity, which is characterized. |
| Interfering Compounds | No false positive or false negative results due to common interfering substances that are not drugs of abuse. | For both Opiates and Oxycodone assays, a large panel of structurally related and unrelated compounds were tested. Generally, non-cross-reactive compounds did not cause false positives at relevant concentrations (-50% CO was NEG). Structurally similar compounds that could cross-react turned POS, as expected for the assay. No tested samples produced a negative result when expected to be positive. |
| Calibrator and Control Stability | Calibrators and controls should be stable for a defined period while maintaining accuracy. | Calibrator stock solution for morphine and oxycodone was stable for one year (within 10% of target value). |
| Storage Stability (Hair Samples) | Drug analytes in hair samples should remain stable over reasonable storage periods. | Opiates stable for 3 years; Oxycodone and Hydrocodone stable for 2 years. Mean change in concentration ranged from -5% to +7% for various analytes. |
| Shipping Stability | Temperature and humidity variations during shipping should not adversely affect test results. | Average mean % change in screening result prior to and after shipping was 1.9%. Four out of 260 samples showed different screening results but were all near the cutoff (±50%), where variability is expected. |
| Cosmetic Treatment Effects | Acknowledgement and characterization of the impact of cosmetic treatments on drug detection. The ELISA protocol itself should not introduce new artifacts related to treatment. | Cosmetic treatments can reduce drug amounts. The study shows the ELISA protocol is not an exception. Bleach, Dye, Permanent, Relaxer, and Shampoo treatments all showed varying effects, mostly decreasing concentrations. Changes at cutoff were observed (e.g., Dyeing Opiates: 57 Pos to Neg, 110 Neg to Pos due to change at cutoff level; Shampoo Oxycodone/Hydrocodone: Pos to Neg). Cross-reactivity was noted for specific treatments (e.g., Permanent with Hydrocodone to Opiates). |
| Environmental Contamination | The assay should be able to distinguish between true positive samples and those with external contamination, facilitated by a methanol wash. | Analytically negative samples exposed to drugs remained negative after methanol wash. Clinically positive samples remained positive after methanol wash. This indicates the methanol wash procedure mitigates false positives from external contamination. |
2. Sample Sizes Used for the Test Set and Data Provenance
-
Test Set Sample Sizes:
- Intra-assay Precision: 11 replicates per concentration level (for Opiates & Oxycodone), 10 replicates per concentration level (for Hydrocodone).
- Inter-assay Precision: 220 samples per concentration level (for Opiates & Oxycodone), 100 samples per concentration level (for Hydrocodone) tested over 20 non-consecutive days.
- Agreement Studies (with GC/MS):
- Opiates: 226 head and body hair samples (176 head hair, 50 body hair).
- Oxycodone & Hydrocodone: 530 head and body hair samples (240 head hair in Study 1, 240 head hair in Retrospective Study 2, 50 body hair in Study 3). Note: Actual N for Oxycodone was 483 (due to exclusions) and for Hydrocodone was 500 (due to exclusions).
- Cross-reactivity: 3 replicates per compound, per concentration level.
- Interfering Compounds: 3 replicates per compound, per concentration level (-50% CO, +125% CO, +150% CO).
- Storage Stability: 54 samples varying in ethnic origin, hair color, and curvature.
- Shipping Study: 260 head hair samples (155 confirmed positive, 100 screened negative, 5 below cutoff).
- Cosmetic Treatment: 176 hair specimens (112 positive for opiates/oxycodone, 64 negative).
- Environmental Contamination: A set of drug-free hair samples and a set of known positive samples. (Exact numbers not specified, but implied to be sufficient for the qualitative determination described).
-
Data Provenance: The document does not explicitly state the country of origin for the human hair samples used in the test sets. It mentions "body and head hair samples from different ages, gender, ethnicities and hair color," but no geographical origin.
-
Retrospective/Prospective: The agreement studies included a "retrospective analysis Study 2" for Oxycodone and Hydrocodone, implying some of the data was collected previously. Other studies (Precision, Study 1 for Agreement, Storage, Shipping, etc.) appear to be prospectively conducted for this submission, although this is not explicitly stated.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications
The ground truth for the test set (agreement studies) was established using Gas Chromatography/Mass Spectrometry (GC/MS) operating in the selected ion monitoring (SIM) mode as the preferred method, with deuterated internal standards. This is an instrumental, objective method, not dependent on human expert interpretation in the same way an imaging study would be. Therefore, the concept of "number of experts" and their "qualifications" for establishing ground truth doesn't directly apply in this context. The validity of GC/MS as a "more specific alternate chemical method" implies its established status as a gold standard in drug testing.
4. Adjudication Method for the Test Set
Since the ground truth for the test set was established by GC/MS, an instrumental method, human adjudication methods like "2+1" or "3+1" are not applicable. The GC/MS results serve as the definitive reference. Discordant results between the ELISA and GC/MS were reported and analyzed, but not "adjudicated" by human experts in the sense of consensus building for ground truth.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
This is an in vitro diagnostic (IVD) hair drug screening assay, not an AI-powered diagnostic imaging device. Therefore, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study focusing on human reader improvement with AI assistance is not relevant and was not performed.
6. If a Standalone (i.e. algorithm only without human-in-the loop performance) was done
The device is an ELISA assay, which is a laboratory-based, instrumental test. The results are generated by the assay (the "algorithm" in a chemical sense) detecting the presence of analytes above a cutoff. The "human-in-the-loop" aspect exists in performing the lab procedures, running the instrument, and interpreting the qualitative (positive/negative) output based on the cutoff. The performance studies (precision, agreement) inherently reflect the standalone performance of the assay. There's no separate "human-in-the-loop" performance that would be contrasted with pure "algorithm only" performance beyond the standard lab process.
7. The Type of Ground Truth Used (expert consensus, pathology, outcomes data, etc.)
The primary ground truth used for agreement studies was Gas Chromatography/Mass Spectrometry (GC/MS) operating in the selected ion monitoring (SIM) mode. This is a highly specific and sensitive analytical method widely considered a confirmatory or gold standard in toxicology for identifying and quantifying substances.
8. The Sample Size for the Training Set
The document does not explicitly mention a "training set" in the context of an algorithm or AI model development. This device is an immunoassay (ELISA), which is an analytical chemical test. Its performance is based on the specificity and reactivity of antibodies and other chemical components, not on a machine learning model trained on data. Therefore, the concept of a "training set" as understood in AI/ML is not applicable here. The development and optimization of the assay would typically involve extensive R&D and validation steps, but these are not referred to as "training" in the AI sense.
9. How the Ground Truth for the Training Set Was Established
As explained in point 8, the concept of a "training set" for an AI/ML algorithm does not apply to this ELISA-based in vitro diagnostic device. The ground truth for the validation/performance studies was established by GC/MS, as detailed in point 7.
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Image /page/0/Picture/1 description: The image shows the seal of the Department of Health & Human Services - USA. The seal is circular, with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. In the center of the seal is an abstract image of an eagle.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
January 8, 2015
OMEGA LABORATORIES, INC. ROBERT BARD MANAGING DIRECTOR 400 NORTH CLEVELAND MOGADORE OH 44260
Re: K140671
Trade/Device Name: Omega Laboratories Hair Drug Screening Assay, (Opiates, Oxycodone and Hydrocodone)
Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate test system Regulatory Class: II Product Code: DJG Dated: January 5, 2015 Received: January 6, 2015
Dear Mr. Robert Bard:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
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You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours,
Stayce Beck -S
For : Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K140671
Device Name
Omega Laboratories Inc. Hair Drug Screening Assay (Opiates, Oxycodone and Hydrocodone)
Indications for Use (Describe)
The Omega Laboratories Hair Drug Screening Assay (Opiates, Oxycodone and Hydrocodone) is an in vitro diagnostic test that is intended for the qualitative detection of opiates (calibrated with morphine) and oxycodone and hydrocodone (calibrated with oxycodone) at or above 300 pg/mg in human head and body hair. To confirm a screen positive result. a more specific alternate chemical method, such as Gas ChromatographyMass Spectrometry (GC/MS) operating in the selected ion monitoring (SIM) mode is the preferred method with deuterated internal standards. Professional judgment should be applied to any drug of abuse test result, particularly when presumptive positive results are obtained.
This test is intended exclusively for single laboratory use only and is not intended for sale to anyone.
| Type of Use (Select one or both, as applicable) |
|---|
| Prescription Use (Part 21 CFR 801 Subpart D) |
| Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) SUMMARY
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92
| 510(k) Number: | K140671 |
|---|---|
| Date of Summary: | January 5, 2015 |
| Applicant: | William R. CorlChief Executive OfficerOmega Laboratories, Inc.400 North ClevelandMogadore, OH 44260Tel: 330-628-5748Fax: 330-628-5803 |
| Correspondent:Name: | ROBERT J BARD, JD, CQE, RAC |
| Address: | Omega Laboratories400 North Cleveland, Mogadore, OH 44260 |
| Phone Number: | 248-573-5040 |
| rbard@reglaw.net | |
| Product Name:Trade Name: | Omega Laboratories Inc. Hair Drug Screening Assay (Opiates, Oxycodone and Hydrocodone) |
| Common Name: | Hair Drug Screening Assay Opiates |
| Regulation Number: | CFR 862.3650 (ProCode DJG) |
| Classification Name: | Opiate test system. |
| Classification Panel: | 91 (Toxicology) |
| Predicate Device: | Omega Hair Drug Screening Assay for Opiates, Oxycodone and Hydrocodone (K103161) |
| Indication for Use: | The Omega Laboratories Hair Drug Screening Assay (Opiates, Oxycodone and Hydrocodone) is an in vitro diagnostic test that is intended for the qualitative detection of opiates (calibrated with morphine) and oxycodone and hydrocodone (calibrated with oxycodone) at or above 300 pg/mg in human head and body hair. To confirm a screen positive result, a more specific alternate chemica method, such as Gas Chromatography/Mass Spectrometry (GC/MS) operating in the selected ion monitoring (SIM) mode is the preferrec method with deuterated internal standards. Professional judgment should be applied to any drug of abuse test result, particularly when presumptive positive results are obtained.This test is intended exclusively for single laboratory use only and is not intended for sale to anyone |
| Comparison: | When used to qualitatively detect Opiates, Oxycodone andHydrocodone in head and body hair specimens collected with theOmega Specimen Collection Device, the Omega assays yield results insubstantial agreement with the predicate device (see Table 1 below). |
| Comparison PerformanceData: | All performance studies demonstrated that the Omega assay is insubstantial agreement with the predicate products.Results obtained from donor specimens showed that the qualitativeresults from the new assays are substantially equivalent to thoseobtained from the predicate devices. |
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Page 2 of 33
Table 1: Comparison of Omega Hair Drug Screening Assay for Opiates, Oxycodone and Hydrocodone
| ComparisonElement -Similarities | Hair Drug Screening Assay for Opiates,Oxycodone and Hydrocodone. (Subjectdevices K140671) | Hair Drug Screening Assay Opiates,Oxycodone and Hydrocodone (Predicatedevice K103161) |
|---|---|---|
| Laboratory | Omega Laboratories, Inc. | Same. |
| Indication for/Intended Use | The Omega Laboratories Hair DrugScreening Assay (Opiates, Oxycodoneand Hydrocodone) is an in vitrodiagnostic test that is intended for thequalitative detection of opiates (calibratedwith morphine) and oxycodone andhydrocodone (calibrated with oxycodone)at or above 300 pg/mg in human headand body hair. To confirm a screenpositive result, a more specific alternatechemical method, such as GasChromatography/Mass Spectrometry(GC/MS) operating in the selected ionmonitoring (SIM) mode is the preferredmethod with deuterated internal standards.Professional judgment should be appliedto any drug of abuse test result,particularly when presumptive positiveresults are obtained.This test is intended exclusively for singlelaboratory use only and is not intended forsale to anyone. | The Omega Laboratories Hair DrugScreening Assays are test systems thatutilize ELISA assays for the qualitativedetection of morphine and related opiates(calibrated with morphine) and oxycodoneand hydrocodone (calibrated withoxycodone) at or above 300 pg/mg in headhair samples.The Omega Laboratories Hair DrugScreening Assay for Opiates, Oxycodoneand Hydrocodone provide only preliminaryanalytical test results. A more specificalternate chemical method must be used inorder to obtain a confirmed result. GasChromatograph - Mass Spectrometryoperating in the selected ion monitoring(SIM) mode or GC/MS/MS in selectedreaction mode (SRM) is the preferredmethod with deuterated internal standards. |
| Method ofMeasurement | Microplate reader. Read at 450 nm | Same. |
| Matrix | Head and body hair | Head hair |
| Cutoffconcentration | 300 pg Opiates, Oxycodone and Hydrocodone/mg hair | Same. |
| Assay Principal | ELISA | Same. |
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Page 3 of 33
Table 1: Comparison of Omega Hair Drug Screening Assay for Opiates, Oxycodone and Hydrocodone
| ComparisonElement -Similarities | Hair Drug Screening Assay for Opiates,Oxycodone and Hydrocodone. (Subjectdevices K140671) | Hair Drug Screening Assay Opiates,Oxycodone and Hydrocodone (Predicatedevice K103161) |
|---|---|---|
| ExtractionMethod | Utilized acid-methanol vs. methanol aloneto facilitate extraction of drug from hair.Proprietary and patent pending method ofpulverizing hair vs. cutting the hair intosmall segments prior to acid-methanolextraction. This improved extraction timeswithout loss of efficiency | Same. |
Performance Studies
PRECISION :
Intra-assay precision studies were performed using 11 replicates of negative head hair samples spiked to the following concentrations of opiates, oxycodone; and hydrocodone: zero drug, -75%, -50%, -25% below the cutoff, and +25%, +50%, +75% and+100% above the cutoff. All samples were treated and analyzed in the same manner as donor hair samples and measured in one run. Head hair was used in this study.
| Drug | Concentration ofSample (pg/mg) | Number ofReplicates | Results #Negative | Results #Positive |
|---|---|---|---|---|
| Opiates | 0 | 11 | 11 | 0 |
| Opiates | 75 | 11 | 11 | 0 |
| Opiates | 150 | 11 | 11 | 0 |
| Opiates | 225 | 11 | 11 | 0 |
| Opiates | 375 | 11 | 0 | 11 |
| Opiates | 450 | 11 | 0 | 11 |
| Opiates | 525 | 11 | 0 | 11 |
| Opiates | 600 | 11 | 0 | 11 |
| Oxycodone | 0 | 11 | 11 | 0 |
| Oxycodone | 75 | 11 | 11 | 0 |
| Oxycodone | 150 | 11 | 11 | 0 |
| Oxycodone | 225 | 11 | 11 | 0 |
| Oxycodone | 375 | 11 | 0 | 11 |
| Oxycodone | 450 | 11 | 0 | 11 |
| Oxycodone | 525 | 11 | 0 | 11 |
| Oxycodone | 600 | 11 | 0 | 11 |
Table 2: Intra-Assay Precision Studies (opiates, oxycodone and hydrocodone)
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Page 4 of 33
| Drug | Concentration ofSample (pg/mg) | Number ofReplicates | Results #Negative | Results #Positive |
|---|---|---|---|---|
| Hydrocodone | 0 | 10 | 10 | 0 |
| Hydrocodone | 75 | 10 | 10 | 0 |
| Hydrocodone | 150 | 10 | 10 | 0 |
| Hydrocodone | 225 | 10 | 9 | 1 |
| Hydrocodone | 375 | 10 | 0 | 10 |
| Hydrocodone | 450 | 10 | 0 | 10 |
| Hydrocodone | 525 | 10 | 0 | 10 |
| Hydrocodone | 600 | 10 | 0 | 10 |
Table 2: Intra-Assay Precision Studies (opiates, oxycodone and hydrocodone)
Inter-assay precision studies were performed using negative hair samples spiked to the following concentrations of opiates, oxycodone and hydrocodone: zero drug, -75%, -25% below the cutoff, and +25%, +50%, +75% and+100% above the cutoff.
All samples were treated and analyzed in the same manner as donor hair samples, which is summarized in Section 9.0. Eleven replicates of these were prepared and analyzed over 20 non-consecutive days. The results of this study are summarized in the tables below and the raw data is attached.
Tables 15a through 15c summarize the result of the Opiates, Oxycodone and Hydrocodone Inter-Assay Precision testing of the Omega Laboratories Drug Screening in Hair Assays.
| Drug | Conc. (pg/mg)(% of Cutoff) | NumberTested | Negative | Positive |
|---|---|---|---|---|
| Opiates | 0 (Negative) | 220 | 220 | 0 |
| Opiates | 75 (-75) | 220 | 220 | 0 |
| Opiates | 150 (-50) | 220 | 220 | 0 |
| Opiates | 225 (-25) | 220 | 220 | 0 |
| Opiates | 375 (125) | 220 | 0 | 220 |
| Opiates | 450 (150) | 220 | 0 | 220 |
| Opiates | 525 (175) | 220 | 0 | 220 |
| Opiates | 600 (200) | 220 | 0 | 220 |
Table 3a: Inter-assay Opiates Precision Study Summary (non-normalized)
Table 3b: Inter-assay Oxycodone Precision Study Summary (non-normalized)
| Drug | Conc. (pg/mg)(% of Cutoff) | NumberTested | Negative | Positive |
|---|---|---|---|---|
| Oxycodone | 0 (Negative) | 220 | 220 | 0 |
| Oxycodone | 75 (-75) | 220 | 220 | 0 |
| Oxycodone | 150 (-50) | 220 | 220 | 0 |
| Oxycodone | 225 (-25) | 220 | 220 | 0 |
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| Drug | Conc. (pg/mg)(% of Cutoff) | NumberTested | Negative | Positive |
|---|---|---|---|---|
| Oxycodone | 375 (125) | 220 | 0 | 220 |
| Oxycodone | 450 (150) | 220 | 0 | 220 |
| Oxycodone | 525 (175) | 220 | 0 | 220 |
| Oxycodone | 600 (200) | 220 | 0 | 220 |
Table 3c: Inter-assay Hydrocodone Precision Study Summary (non-normalized
| Drug | Conc. (pg/mg)% of Cutoff | NumberTested | Negative | Positive |
|---|---|---|---|---|
| Hydrocodone | 0 (Negative) | 100 | 100 | 0 |
| Hydrocodone | 75 (-75) | 100 | 100 | 0 |
| Hydrocodone | 150 (-50) | 100 | 100 | 0 |
| Hydrocodone | 225 (-25) | 100 | 61 | 39 |
| Hydrocodone | 375 (125) | 100 | 0 | 100 |
| Hydrocodone | 450 (150) | 100 | 0 | 100 |
| Hydrocodone | 525 (175) | 100 | 0 | 100 |
| Hydrocodone | 600 (200) | 100 | 0 | 100 |
AGREEMENT STUDIES:
The method comparison was performed using two opiates studies by testing 226 head and body hair samples consisting of 176 head hair samples in Study 1 and 50 body hair samples in Study 3.
Agreement studies also included 530 head and body hair samples that were tested in three oxycodone and hydrocodone studies consisting of 240 head hair samples in Study 1, 240 head hairs samples in retrospective analysis Study 2 and 50 body hair samples in Study 3.
In the studies, each specimen was divided into two aliquots and one was used for screening and the other for GC/MS confirmation. Testing was performed on body and head hair samples from different ages, gender, ethnicities and hair color. The results were:
Opiates Agreement Studies (from Studies 1 and 3)
| Table 3a: Opiates Equivalents Summary of Agreement Study Results (n=226) Head and Body | ||
|---|---|---|
| -- | -- | ---------------------------------------------------------------------------------------- |
| ELISA TestResult | Negative byGC/MS | Less than halfthe cutoffconcentrationby GC/MS | Near CutoffNegative(Between 50%below thecutoff and thecutoffconcentration) | Near CutoffPositive(Between thecutoff and 50%above the cutoffconcentration) | High Positive(Greater that50% above thecutoffconcentration) |
|---|---|---|---|---|---|
| Positive | 0 | 0 | 2 | 24 | 116 |
| Negative | 70 | 4 | 9 | 1 | 0 |
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| SampleNo. | Screening Cutoff(pg/mg) | ELISA TestResult(POS/NEG) | GC/MS Cutoff(pg/mg) | GC/MS Drug Result (pg/mg) |
|---|---|---|---|---|
| 16 | 300 | POS | 300 | HDC 268 |
| 150 | 300 | POS | 300 | HDC 299 |
| 29 | 300 | NEG | 300 | HDC 354 |
Table 3b: GC/MS Summary of Opiates Equivalents Discordant Results
Oxycodone and Hydrocodone Agreement Studies (from Studies 1,2 and 3)
| ELISATestResult | Negative byGC/MS | Less than halfthe cutoffconcentrationby GC/MS | Near CutoffNegative(Between 50%below thecutoff and thecutoffconcentration) | Near CutoffPositive(Between thecutoff and 50%above the cutoffconcentration) | High Positive(Greater that50% abovethe cutoffconcentration) |
|---|---|---|---|---|---|
| Positive | 0 | 0 | 6 | 94 | 221 |
| Negative | 140 | 6 | 14 | 2 | 0 |
*Of the 530 samples compared in the Oxy assays, 46 ELISA positive head hair samples were negative by GC/MS confirmation for oxycodone but positive for hydrocodone and one head hair test sample was lost during centrifugation reducing the number of test samples reported here to 483.
| Table 4b: GC/MS Summary of Oxycodone Discordant Results | |||
|---|---|---|---|
| -- | -- | --------------------------------------------------------- | -- |
| SampleNo. | Screening Cutoff(pg/mg) | ELISA TestResult(POS/NEG) | GC/MS Cutoff(pg/mg) | GC/MS Drug Result (pg/mg) |
|---|---|---|---|---|
| 835334 | 300 | NEG | 300 | OXY 169HDC 167 |
| EXOP 39 | 300 | POS | 300 | OXY 131HDC 83 |
| 788840 | 300 | POS | 300 | OXY 185HDC 29 |
| 787180c | 300 | POS | 300 | OXY 229HDC 68 |
| 789526 | 300 | POS | 300 | OXY 240HDC 49 |
| 872104 | 300 | NEG | 300 | OXY 130HDC 221 |
{9}------------------------------------------------
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| Table 4b: GC/MS Summary of Oxycodone Discordant Results | |
|---|---|
| SampleNo. | Screening Cutoff(pg/mg) | ELISA TestResult(POS/NEG) | GC/MS Cutoff(pg/mg) | GC/MS Drug Result (pg/mg) |
|---|---|---|---|---|
| 854976 | 300 | POS | 300 | OXY 255 |
| 800488 | 300 | POS | 300 | OXY 298 |
| Table 4c: Hydrocodone Summary of Agreement Study Results (n=500*) | |
|---|---|
| ELISATestResult | Negative byGC/MS | Less than halfthe cutoffconcentrationby GC/MS | Near CutoffNegative(Between 50%below thecutoff and thecutoffconcentration) | Near CutoffPositive(Between thecutoff and 50%above the cutoffconcentration) | High Positive(Greater that50% abovethe cutoffconcentration) |
|---|---|---|---|---|---|
| Positive | 0 | 0 | 8 | 110 | 201 |
| Negative | 142 | 8 | 25 | 6 | 0 |
*Of the 530 samples compared in the HDC assays, 27 ELISA positive head hair samples were negative by GC/MS confirmation for hydrocodone but positive for oxycodone, one head hair test sample was lost during centrifugation and 2 ELISA head hair positive samples were not able to be confirmed by GC/MS which reduced the number of test samples reported here to 500.
| Table 4d: GC/MS Summary of Hydrocodone Discordant Results |
|---|
| ----------------------------------------------------------- |
| SampleNo. | Screening Cutoff(pg/mg) | ELISA TestResult(POS/NEG) | GC/MS Cutoff(pg/mg) | GC/MS Drug Result (pg/mg) |
|---|---|---|---|---|
| 787832 | 300 | NEG | 300 | HDC 403 |
| 89 | 300 | NEG | 300 | HDC 313 |
| 65 | 300 | NEG | 300 | HDC 334 |
| 835334 | 300 | NEG | 300 | HDC 167OXY 169 |
| 872104 | 300 | NEG | 300 | HDC 221OXY 130 |
| 835981a | 300 | NEG | 300 | HDC 347 |
| 788840 | 300 | POS | 300 | HDC 29OXY 185 |
| 789526 | 300 | POS | 300 | HDC 49OXY 240 |
| 787180c | 300 | POS | 300 | HDC 68OXY 229 |
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| SampleNo. | Screening Cutoff(pg/mg) | ELISA TestResult(POS/NEG) | GC/MS Cutoff(pg/mg) | GC/MS Drug Result (pg/mg) |
|---|---|---|---|---|
| EXOP 39 | 300 | POS | 300 | HDC 83OXY 131 |
| 790194 | 300 | POS | 300 | HDC 204 |
| 849237 | 300 | POS | 300 | HDC 256 |
| 904316 | 300 | POS | 300 | HDC 272 |
| 780598 | 300 | POS | 300 | HDC 283 |
Table 4d: GC/MS Summary of Hydrocodone Discordant Results
CROSSREACTIVITY:
The Cross-reactivity study was designed to evaluate the specificity of the Omega Laboratories, Inc. ELISA Screening Protocol and the possible effect of interfering compounds.
Table 5a: Cross-reactivity of Opiates ELISA with Structurally Similar Compounds:
| Compound | Approximate Concentration of Compound(pg/mg) Equivalent to 300pg/mg OpiatesCutoff Control(n=3) | Percent Cross-reactivity (%) | |
|---|---|---|---|
| Morphine | 300 | 100.0 | |
| Heroin | 200 | 150.0 | |
| Codeine | 250 | 120.0 | |
| 6-Acetylcodeine | 275 | 109.1 | |
| Ethylmorphine | 200 | 150.0 | |
| Dihydrocodeine | 700 | 42.9 | |
| 6-Monoacetylmorphine | 200 | 150.0 | |
| Morphine-3-Beta-Glucuronide | 700 | 42.9 | |
| Thebain | 1250 | 24.0 | |
| Morphine-6-Beta-Glucuronide | 600 | 50.0 | |
| Dihydromorphine | 1500 | 20.0 | |
| Hydrocodone | 1250 | 24.0 | |
| Hydromorphone | 2000 | 15.0 | |
| Nalorphine | 7000 | 4.3 | |
| Levorphanol | 4000 | 7.5 | |
| Norcodeine | 250000 | 0.1 | |
| Oxycodone | 225000 | 0.1 | |
| Normorphine | 175000 | 0.2 | |
| Diprenorphine | 225000 | 0.1 | |
| Dextromethorphan | Not achieved at highest spike concentration. |
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| Compound | Approximate Concentration of Compound(pg/mg) Equivalent to 300pg/mg OpiatesCutoff Control(n=3) | Percent Cross-reactivity (%) |
|---|---|---|
| Naltrexone | Not achieved at highest spike concentration.1000000 pg/mg | |
| Norbuprenorphine | Not achieved at highest spike concentration.1000000 pg/mg | |
| Buprenorphine | Not achieved at highest spike concentration.1000000 pg/mg | |
| Oxymorphone | 200000 | 0.2 |
| Naltriben | Not achieved at highest spike concentration.1000000 pg/mg | |
| Nalmefene | Not achieved at highest spike concentration.1000000 pg/mg | |
| Apomorphine | Not achieved at highest spike concentration.1000000 pg/mg | |
| Naloxone | Not achieved at highest spike concentration.1000000 pg/mg | |
| Noroxymorphone | Not achieved at highest spike concentration.1000000 pg/mg | |
| Noroxycodone | Not achieved at highest spike concentration.1000000 pg/mg | |
| 3-Methoxynaltrexone | Not achieved at highest spike concentration.1000000 pg/mg |
Table 5a: Cross-reactivity of Opiates ELISA with Structurally Similar Compounds:
Table 5b: Cross reactivity of ELISA Oxycodone with Structurally Similar Compounds
| Compound | Approximate Concentration ofCompound (pg/mg) Equivalent to300pg/mg Oxycodone Cutoff Control(n=3) | Percent Cross-reactivity(%) | |
|---|---|---|---|
| Hydrocodone | 250 | 120 | |
| Oxycodone | 300 | 100 | |
| Oxymorphone | 1500 | 20 | |
| Dihydrocodeine | 2500 | 12 | |
| 6-Acetylcodeine | 4000 | 7.5 | |
| Codeine | 4500 | 6.7 | |
| Ethylmorphine | 5000 | 6 | |
| Hydromorphone | 6000 | 5 | |
| Heroin | 15000 | 2 | |
| Dihydromorphine | 15000 | 2 | |
| Levorphanol | 15000 | 2 | |
| Compound | Approximate Concentration ofCompound (pg/mg) Equivalent to300pg/mg Oxycodone Cutoff Control(n=3) | Percent Cross-reactivity(%) | |
| 6-Monoacetylmorphine | 20000 | 1.5 | |
| Morphine | 30000 | 1 | |
| Noroxycodone | 30000 | 1 | |
| Thebaine | 40000 | 0.75 | |
| Morphine-3-β-glucuronide | 150000 | 0.2 | |
| Naloxone | 250000 | 0.12 | |
| Norcodeine | 400000 | 0.07 | |
| Morphine-6-β-glucuronide | Not achieved at highest spike concentration.40000 pg/mg | ||
| Norbuprenorphine | Not achieved at highest spike concentration.40000 pg/mg | ||
| Buprenorphine | Not achieved at highest spike concentration.40000 pg/mg | ||
| Noroxymorphone | Not achieved at highest spike concentration.40000 pg/mg | ||
| Nalorphine | Not achieved at highest spike concentration.40000 pg/mg | ||
| Normorphine | Not achieved at highest spike concentration.400000 pg/mg | ||
| Diprenorphine | Not achieved at highest spike concentration.400000 pg/mg | ||
| Dextromethorphan | Not achieved at highest spike concentration.400000 pg/mg | ||
| Naltrexone | Not achieved at highest spike concentration.400000 pg/mg | ||
| Naltriben | Not achieved at highest spike concentration.400000 pg/mg | ||
| Nalmefene | Not achieved at highest spike concentration.400000 pg/mg | ||
| Apomorphine | Not achieved at highest spike concentration.400000 pg/mg | ||
| 3-Methoxy-naltrexone | Not achieved at highest spike concentration.400000 pg/mg |
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Table 5b: Cross reactivity of ELISA Oxycodone with Structurally Similar Compounds
Effect of Interfering Compounds: A variety of structurally related and unrelated compounds were tested for interference at 10000ng/ml (40000pg/mg) in the Opiates ELISA and the Oxycodone ELISA assays. Negative hair extracts were spiked with morphine or oxycodone at -50% (125pg/mg), +125% (375pg/mg) and +150% (450pg/mg) of the Cutoff Concentration (300pg/mg). These were then additionally spiked with 10000ng/ml (400000pg/mg) of the structurally related compounds unless otherwise noted. The absorbances were compared to the 300 pg/mg Cutoff control (CO). Only compounds that were structurally cross-reactive interfered in the assay. These structurally related compounds produced a
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positive response due to sufficient cross-reactivity. No tested samples produced a negative result when expected to be positive. The analysis was performed in triplicate.
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
|---|---|---|---|
| (-) 11-nor-9-carboxy-delta8-Tetrahydrocannabinol | NEG | POS | POS |
| (-) 11-nor-9-carboxy-delta9-Tetrahydrocannabinol | NEG | POS | POS |
| R (-) Amphetamine | NEG | POS | POS |
| (-) Cotinine | NEG | POS | POS |
| (-) Cotinine -N-oxide | NEG | POS | POS |
| (-) Isoproterenol | NEG | POS | POS |
| (-) Methamphetamine | NEG | POS | POS |
| (-) Nicotine | NEG | POS | POS |
| (-) Phenylephrine | NEG | POS | POS |
| (-)-Alpha-methadol | NEG | POS | POS |
| (+) Amphetamine | NEG | POS | POS |
| (+) Isoproterenol | NEG | POS | POS |
| (+) Methamphetamine | NEG | POS | POS |
| (+) Pseudoephedrine | NEG | POS | POS |
| (±) 11-nor-9-carboxy-delta9-Tetrahydrocannabinol | NEG | POS | POS |
| (±) 2,5-Dimethoxy- 4-bromoamphetamine | NEG | POS | POS |
| (±) Alphaprodine | NEG | POS | POS |
| (±) Ketamine | NEG | POS | POS |
| (±) MBDB | NEG | POS | POS |
| (±) MDA | NEG | POS | POS |
| (±) MDEA | NEG | POS | POS |
| (±) MDMA | NEG | POS | POS |
| (±) Metanephrine | NEG | POS | POS |
| (±) Metoprolol | NEG | POS | POS |
| (±) Norcotinine | NEG | POS | POS |
| (±) Propanolol | NEG | POS | POS |
| (±) Trans-3'-Hydroxycotinine | NEG | POS | POS |
| 11-Hydroxy-delta9-Tetrahydrocannabinol | NEG | POS | POS |
| 19-Nortestosterone (Nandrolone) | NEG | POS | POS |
| 1R,2S (-) Ephedrine | NEG | POS | POS |
| 1S,2R (+) Ephedrine | NEG | POS | POS |
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
| 2-Oxo-3-hydroxy-LSD | NEG | POS | POS |
| 3-Methoxynaltrexone | NEG | POS | POS |
| 3-Trans-Hydroxy-norcotinine | NEG | POS | POS |
| 4-Acetoamidophenol | NEG | POS | POS |
| 4-Hydroxy-Phencyclidine | NEG | POS | POS |
| 5,5-Diphenylhydantoin | NEG | POS | POS |
| 6-Acetyl-codeine | POS | POS | POS |
| 6-MonoacetyImorphine | POS | POS | POS |
| 7-Aminoclonazepam | NEG | POS | POS |
| 7-Aminonitrazepam | NEG | POS | POS |
| Acebutolol | NEG | POS | POS |
| Acetophenetidin | NEG | POS | POS |
| Acetopromazine | NEG | POS | POS |
| Acetylsalicyclic acid | NEG | POS | POS |
| Alfentanil | NEG | POS | POS |
| Alpha-Ergocryptine | NEG | POS | POS |
| Alprazolam | NEG | POS | POS |
| 7-Aminoflunitrazepam | NEG | POS | POS |
| Aminorex | NEG | POS | POS |
| Amitriptyline | NEG | POS | POS |
| Amobarbital | NEG | POS | POS |
| Amoxicillin | NEG | POS | POS |
| Anhydroecgonine | NEG | POS | POS |
| Anileridine | NEG | POS | POS |
| Apomorphine | NEG | POS | POS |
| Atenolol | NEG | POS | POS |
| Azaperone | NEG | POS | POS |
| Benzoylecgonine | NEG | POS | POS |
| Benzoylecgonine isopropyl ester | NEG | POS | POS |
| Betamethasone | NEG | POS | POS |
| Boldenone | NEG | POS | POS |
| Bumetanide | NEG | POS | POS |
| Bupivicaine | NEG | POS | POS |
| Buprenorphine | POS | POS | POS |
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
| Buprenorphine-glucuronide(2500ng/ml) | NEG | POS | POS |
| Buspirone | NEG | POS | POS |
| Butabarbital | NEG | POS | POS |
| Butalbital | NEG | POS | POS |
| Caffeine | NEG | POS | POS |
| Cannabidiol | NEG | POS | POS |
| Cannabinol | NEG | POS | POS |
| Carbamazepine | NEG | POS | POS |
| Carisoprodol | NEG | POS | POS |
| Chlordiazepoxide | NEG | POS | POS |
| Chlorpromazine | NEG | POS | POS |
| Cimeterol | NEG | POS | POS |
| Clenbuterol | NEG | POS | POS |
| Clomipramine | NEG | POS | POS |
| Clonazepam | NEG | POS | POS |
| Clonidine | NEG | POS | POS |
| Clozapine | NEG | POS | POS |
| Cocaethylene | NEG | POS | POS |
| Cocaine | NEG | POS | POS |
| Codeine | POS | POS | POS |
| Corticosterone | NEG | POS | POS |
| Cortisone | NEG | POS | POS |
| Cotinine-N-beta-D-Glucuronide | NEG | POS | POS |
| Cyclobenzaprine | NEG | POS | POS |
| d, I-N-Desmethylvenlafaxine | NEG | POS | POS |
| Delta8-Tetrahydrocannabinol | NEG | POS | POS |
| Delta9-Tetrahydrocannabinol | NEG | POS | POS |
| Deoxycorticosterone | NEG | POS | POS |
| Desalkylflurazepam | NEG | POS | POS |
| Desipramine | NEG | POS | POS |
| Desmethyldoxepin (cis/trans) | NEG | POS | POS |
| Despropionyl-fentanyl | NEG | POS | POS |
| Dexamethasone | NEG | POS | POS |
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
| Dextromethorphan | NEG | POS | POS |
| Diazepam | NEG | POS | POS |
| Dibucaine | NEG | POS | POS |
| Dihydrocodeine | POS | POS | POS |
| Dihydroergotamine | NEG | POS | POS |
| Dihydromorphine | POS | POS | POS |
| Diphenhydramine | NEG | POS | POS |
| Diphenoxylate | NEG | POS | POS |
| Diprenorphine | POS | POS | POS |
| Dothiepin (cis/trans) | NEG | POS | POS |
| Doxepin | NEG | POS | POS |
| Doxylamine | NEG | POS | POS |
| Droperidol | NEG | POS | POS |
| Ecgonine | NEG | POS | POS |
| Ecgonine methyl ester | NEG | POS | POS |
| EDDP | NEG | POS | POS |
| Effexor (Venlafaxine) | NEG | POS | POS |
| EMDP | NEG | POS | POS |
| Ergonovine | NEG | POS | POS |
| Erythromycin | NEG | POS | POS |
| Estazolam | NEG | POS | POS |
| Ethacrynic acid | NEG | POS | POS |
| Ethopropazine | NEG | POS | POS |
| Ethylmorphine | POS | POS | POS |
| Fenfluramine | NEG | POS | POS |
| Fentanyl | NEG | POS | POS |
| Flumethasone | NEG | POS | POS |
| Flunitrazepam | NEG | POS | POS |
| Fluphenazine | NEG | POS | POS |
| Flurazepam | NEG | POS | POS |
| Furosemide | NEG | POS | POS |
| Gentamicin | NEG | POS | POS |
| Gluthimide | NEG | POS | POS |
| Haloperidol | NEG | POS | POS |
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
| Heroin | POS | POS | POS |
| Hexobarbital | NEG | POS | POS |
| HMMA | NEG | POS | POS |
| Hydrochlorothiazide | NEG | POS | POS |
| Hydrocodone | POS | POS | POS |
| Hydrocortisone | NEG | POS | POS |
| Hydromorphone | POS | POS | POS |
| (±) 4-Hydroxyephedrine | NEG | POS | POS |
| Hydroxymethamphetamine | NEG | POS | POS |
| Ibogaine | NEG | POS | POS |
| Ibuprofen | NEG | POS | POS |
| Imipramine | NEG | POS | POS |
| Ketoprofen | NEG | POS | POS |
| LAAM | NEG | POS | POS |
| Labetalol | NEG | POS | POS |
| Levorphanol | POS | POS | POS |
| L-Hyoscyamine | NEG | POS | POS |
| Lidocaine | NEG | POS | POS |
| Lorazepam | NEG | POS | POS |
| LSD | NEG | POS | POS |
| Lysergic acid | NEG | POS | POS |
| Lysergol | NEG | POS | POS |
| Maprotiline | NEG | POS | POS |
| Meperidine | NEG | POS | POS |
| Mephentermine | NEG | POS | POS |
| Mepivacaine | NEG | POS | POS |
| Metaphit | NEG | POS | POS |
| Metaproterenol | NEG | POS | POS |
| Metaraminol | NEG | POS | POS |
| Methadone | NEG | POS | POS |
| Methohexital | NEG | POS | POS |
| Methoxyphenamine | NEG | POS | POS |
| Methylergonovine | NEG | POS | POS |
| Methylphenidate | NEG | POS | POS |
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
| m-Hydroxybenzoylecgonine | NEG | POS | POS |
| Mianserin | NEG | POS | POS |
| Midazolam | NEG | POS | POS |
| Monensin | NEG | POS | POS |
| Morphine | POS | POS | POS |
| Morphine-3-betaglucuronide | POS | POS | POS |
| Morphine-6-betaglucuronide | POS | POS | POS |
| Nadolol | NEG | POS | POS |
| Nalmefene | NEG | POS | POS |
| Nalorphine | POS | POS | POS |
| Naloxone-3-beta-D-glucuronide | NEG | POS | POS |
| Naltrexone | NEG | POS | POS |
| Naltriben | NEG | POS | POS |
| Naproxen | NEG | POS | POS |
| N-Desmethylclomipramine | NEG | POS | POS |
| N-Desmethylflunitrazepam | NEG | POS | POS |
| N-Desmethyltramadol | NEG | POS | POS |
| N-Desmethyltrimipramine | NEG | POS | POS |
| Neomycin | NEG | POS | POS |
| Nitrazepam | NEG | POS | POS |
| Norbenzoylecgonine | NEG | POS | POS |
| Norbuprenorphine | NEG | POS | POS |
| Norcocaethylene | NEG | POS | POS |
| Norcocaine | NEG | POS | POS |
| Norcodeine | POS | POS | POS |
| Nordiazepam | NEG | POS | POS |
| Norfentanyl | NEG | POS | POS |
| Nor-LAAM | NEG | POS | POS |
| Nor-LSD/Nor-ISO-LSD | NEG | POS | POS |
| Normeperidine | NEG | POS | POS |
| Normeperidinic acid | NEG | POS | POS |
| Normorphine | POS | POS | POS |
| Noroxycodone | NEG | POS | POS |
| Noroxymorphone | POS | POS | POS |
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
| Norpropoxyphene | NEG | POS | POS |
| Nortriptyline | NEG | POS | POS |
| Noscapine | NEG | POS | POS |
| O-Desmethyltramadol | NEG | POS | POS |
| Oxazepam | NEG | POS | POS |
| Oxprenolol | NEG | POS | POS |
| Oxycodone | POS | POS | POS |
| Oxymorphone | POS | POS | POS |
| p-Acetamidophenyl-beta-D-glucuronide | NEG | POS | POS |
| Papaverine | NEG | POS | POS |
| Pemoline | NEG | POS | POS |
| Penicillin G | NEG | POS | POS |
| Pentazocine | NEG | POS | POS |
| Pentobarbital | NEG | POS | POS |
| Perphenazine | NEG | POS | POS |
| Phendimetrazine | NEG | POS | POS |
| Phenelzine | NEG | POS | POS |
| Phenobarbital | NEG | POS | POS |
| Phenothiazine | NEG | POS | POS |
| Phentermine | NEG | POS | POS |
| Phenylbutazone | NEG | POS | POS |
| Phenylethyamine | NEG | POS | POS |
| Phenylpropanolamine | NEG | POS | POS |
| PMA | NEG | POS | POS |
| PMMA | NEG | POS | POS |
| Prednisolone | NEG | POS | POS |
| Prilocaine | NEG | POS | POS |
| Prochlorperazine | NEG | POS | POS |
| Progesterone | NEG | POS | POS |
| Promazine | NEG | POS | POS |
| Promethazine | NEG | POS | POS |
| Propiomazine | NEG | POS | POS |
| Propionylpromazine | NEG | POS | POS |
| Propoxyphene | NEG | POS | POS |
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
| Protriptyline | NEG | POS | POS |
| Quinidine | NEG | POS | POS |
| R (+) Methcathinone | NEG | POS | POS |
| R (-) Epinephrine | NEG | POS | POS |
| R (+) Cathinone | NEG | POS | POS |
| Salbutamol | NEG | POS | POS |
| Secobarbital | NEG | POS | POS |
| Sertraline | NEG | POS | POS |
| Stanazalol | NEG | POS | POS |
| Streptomycin | NEG | POS | POS |
| Sulfadimethoxine | NEG | POS | POS |
| Sulfamethazine | NEG | POS | POS |
| Sulfathiazole | NEG | POS | POS |
| Temazepam | NEG | POS | POS |
| Terbutaline | NEG | POS | POS |
| Tetracycline | NEG | POS | POS |
| Thebaine | POS | POS | POS |
| Theophylline | NEG | POS | POS |
| Thioridazine | NEG | POS | POS |
| Tramadol | NEG | POS | POS |
| Triamcinolone | NEG | POS | POS |
| Triazolam | NEG | POS | POS |
| Trifluoperazine | NEG | POS | POS |
| Trifluopromazine | NEG | POS | POS |
| Trimeprazine | NEG | POS | POS |
| Trimipramine | NEG | POS | POS |
| Tylosin | NEG | POS | POS |
| Tyramine | NEG | POS | POS |
| Yohimbic acid | NEG | POS | POS |
| Yohimbine | NEG | POS | POS |
| Zolpidem | NEG | POS | POS |
| Zopiclone | NEG | POS | POS |
| Phencyclidine | NEG | POS | POS |
| R,R (-)-Pseudoephedrine | NEG | POS | POS |
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
| Phencyclidine Morpholine | NEG | POS | POS |
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Table 5c: Interferences of Structurally Related and Unrelated Compounds on Opiates ELISA Assay
Table 5d: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Sys
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
|---|---|---|---|
| (-) 11-nor-9-carboxy-delta8-Tetrahydrocannabinol | NEG | POS | POS |
| (-) 11-nor-9-carboxy-delta9-Tetrahydrocannabinol | NEG | POS | POS |
| R (-) Amphetamine | NEG | POS | POS |
| (-) Cotinine | NEG | POS | POS |
| (-) Cotinine -N-oxide | NEG | POS | POS |
| (-) Isoproterenol | NEG | POS | POS |
| (-) Methamphetamine | NEG | POS | POS |
| (-) Nicotine | NEG | POS | POS |
| (-) Phenylephrine | NEG | POS | POS |
| (-)-Alpha-methadol | NEG | POS | POS |
| (+) Amphetamine | NEG | POS | POS |
| (+) Isoproterenol | NEG | POS | POS |
| (+) Methamphetamine | NEG | POS | POS |
| (+) Pseudoephedrine | NEG | POS | POS |
| (±) 11-nor-9-carboxy-delta9-Tetrahydrocannabinol | NEG | POS | POS |
| (±) 2,5-Dimethoxy- 4-bromoamphetamine | NEG | POS | POS |
| (±) Alphaprodine | NEG | POS | POS |
| (±) Ketamine | NEG | POS | POS |
| (±) MBDB | NEG | POS | POS |
| (±) MDA | NEG | POS | POS |
| (±) MDEA | NEG | POS | POS |
| (±) MDMA | NEG | POS | POS |
| (±) Metanephrine | NEG | POS | POS |
| (±) Metoprolol | NEG | POS | POS |
| (±) Norcotinine | NEG | POS | POS |
| (±) Propanolol | NEG | POS | POS |
| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
| (±) Trans-3'-Hydroxycotinine | NEG | POS | POS |
| 11-Hydroxy-delta9-Tetrahydrocannabinol | NEG | POS | POS |
| 19-Nortestosterone (Nandrolone) | NEG | POS | POS |
| 1R,2S (-) Ephedrine | NEG | POS | POS |
| 1S,2R (+) Ephedrine | NEG | POS | POS |
| 2-Oxo-3-hydroxy-LSD | NEG | POS | POS |
| 3-Methoxynaltrexone | POS | POS | POS |
| 3-Trans-Hydroxy-norcotinine | NEG | POS | POS |
| 4-Acetoamidophenol | NEG | POS | POS |
| 4-Hydroxy-Phencyclidine | NEG | POS | POS |
| 5,5-Diphenylhydantoin | NEG | POS | POS |
| 6-Acetyl-codeine | POS | POS | POS |
| 6-MonoacetyImorphine | POS | POS | POS |
| 7-Aminoclonazepam | NEG | POS | POS |
| 7-Aminonitrazepam | NEG | POS | POS |
| Acebutolol | NEG | POS | POS |
| Acetophenetidin | NEG | POS | POS |
| Acetopromazine | NEG | POS | POS |
| AcetyIsalicyclic acid | NEG | POS | POS |
| Alfentanil | NEG | POS | POS |
| Alpha-Ergocryptine | NEG | POS | POS |
| Alprazolam | NEG | POS | POS |
| 7-Aminoflunitrazepam | NEG | POS | POS |
| Aminorex | NEG | POS | POS |
| Amitriptyline | NEG | POS | POS |
| Amobarbital | NEG | POS | POS |
| Amoxicillin | NEG | POS | POS |
| Anhydroecgonine | NEG | POS | POS |
| Anileridine | NEG | POS | POS |
| Apomorphine | NEG | POS | POS |
| Atenolol | NEG | POS | POS |
| Atropine | NEG | POS | POS |
| Azaperone | NEG | POS | POS |
| Benzoylecgonine | NEG | POS | POS |
{22}------------------------------------------------
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Table 5d: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Sys
{23}------------------------------------------------
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| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
|---|---|---|---|
| Benzoylecgonine isopropyl ester | NEG | POS | POS |
| Betamethasone | NEG | POS | POS |
| Boldenone | NEG | POS | POS |
| Bumetanide | NEG | POS | POS |
| Bupivicaine | NEG | POS | POS |
| Buprenorphine | NEG | POS | POS |
| Buspirone | NEG | POS | POS |
| Butabarbital | NEG | POS | POS |
| Butalbital | NEG | POS | POS |
| Caffeine | NEG | POS | POS |
| Cannabidiol | NEG | POS | POS |
| Cannabinol | NEG | POS | POS |
| Carbamazepine | NEG | POS | POS |
| Carisoprodol | NEG | POS | POS |
| Chlordiazepoxide | NEG | POS | POS |
| Chlorpromazine | NEG | POS | POS |
| Cimeterol | NEG | POS | POS |
| Clenbuterol | NEG | POS | POS |
| Clomipramine | NEG | POS | POS |
| Clonazepam | NEG | POS | POS |
| Clonidine | NEG | POS | POS |
| Clozapine | NEG | POS | POS |
| Cocaethylene | NEG | POS | POS |
| Cocaine | NEG | POS | POS |
| Codeine | POS | POS | POS |
| Corticosterone | NEG | POS | POS |
| Cortisone | NEG | POS | POS |
| Cotinine-N-beta-D-Glucuronide | NEG | POS | POS |
| Cyclobenzaprine | NEG | POS | POS |
| d,I-N-Desmethylvenlafaxine | NEG | POS | POS |
| Delta8-Tetrahydrocannabinol | NEG | POS | POS |
| Delta9-Tetrahydrocannabinol | NEG | POS | POS |
| Deoxycorticosterone | NEG | POS | POS |
| Desalkylflurazepam | NEC | POS | POS |
Table 5d: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Sys
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| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
|---|---|---|---|
| Desipramine | NEG | POS | POS |
| Desmethyldoxepin (cis/trans) | NEG | POS | POS |
| Despropionyl-fentanyl | NEG | POS | POS |
| Dexamethasone | NEG | POS | POS |
| Dextromethorphan | NEG | POS | POS |
| Diazepam | NEG | POS | POS |
| Dibucaine | NEG | POS | POS |
| Dihydrocodeine | POS | POS | POS |
| Dihydroergotamine | NEG | POS | POS |
| Dihydromorphine | POS | POS | POS |
| Diphenhydramine | NEG | POS | POS |
| Diphenoxylate | NEG | POS | POS |
| Diprenorphine | NEG | POS | POS |
| Dothiepin (cis/trans) | NEG | POS | POS |
| Doxepin | NEG | POS | POS |
| Doxylamine | NEG | POS | POS |
| Droperidol | NEG | POS | POS |
| Ecgonine | NEG | POS | POS |
| Ecgonine methyl ester | NEG | POS | POS |
| EDDP | NEG | POS | POS |
| Effexor (Venlafaxine) | NEG | POS | POS |
| EMDP | NEG | POS | POS |
| Ergonovine | NEG | POS | POS |
| Erythromycin | NEG | POS | POS |
| Estazolam | NEG | POS | POS |
| Ethacrynic acid | NEG | POS | POS |
| Ethopropazine | NEG | POS | POS |
| Ethylmorphine | POS | POS | POS |
| Fenfluramine | NEG | POS | POS |
| Fentanyl | NEG | POS | POS |
| Flumethasone | NEG | POS | POS |
| Flunitrazepam | NEG | POS | POS |
| Fluphenazine | NEG | POS | POS |
| Flurazepam | NEG | POS | POS |
Table 5d: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Sys
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| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
|---|---|---|---|
| Furosemide | NEG | POS | POS |
| Gentamicin | NEG | POS | POS |
| Gluthimide | NEG | POS | POS |
| Haloperidol | NEG | POS | POS |
| Heroin | POS | POS | POS |
| Hexobarbital | NEG | POS | POS |
| HMMA | NEG | POS | POS |
| Hydrochlorothiazide | NEG | POS | POS |
| Hydrocodone | POS | POS | POS |
| Hydrocortisone | NEG | POS | POS |
| Hydromorphone | POS | POS | POS |
| (±) 4-Hydroxyephedrine | NEG | POS | POS |
| Hydroxymethamphetamine | NEG | POS | POS |
| Ibogaine | NEG | POS | POS |
| Ibuprofen | NEG | POS | POS |
| Imipramine | NEG | POS | POS |
| Ketoprofen | NEG | POS | POS |
| LAAM | NEG | POS | POS |
| Labetalol | NEG | POS | POS |
| LAMPA (1000ng/ml) | NEG | POS | POS |
| Levorphanol | POS | POS | POS |
| L-Hyoscyamine | NEG | POS | POS |
| Lidocaine | NEG | POS | POS |
| Lorazepam | NEG | POS | POS |
| LSD | NEG | POS | POS |
| Lysergic acid | NEG | POS | POS |
| Lysergol | NEG | POS | POS |
| Maprotiline | NEG | POS | POS |
| Meperidine | NEG | POS | POS |
| Mephentermine | NEG | POS | POS |
| Mepivacaine | NEG | POS | POS |
| Metaphit | NEG | POS | POS |
| Metaproterenol | NEG | POS | POS |
| Metaraminol | NEG | POS | POS |
Table 5d: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Sys
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| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
|---|---|---|---|
| Methadone | NEG | POS | POS |
| Methohexital | NEG | POS | POS |
| Methoxyphenamine | NEG | POS | POS |
| Methylergonovine | NEG | POS | POS |
| Methylphenidate | NEG | POS | POS |
| m-Hydroxybenzoylecgonine | NEG | POS | POS |
| Mianserin | NEG | POS | POS |
| Midazolam | NEG | POS | POS |
| Monensin | NEG | POS | POS |
| Morphine | POS | POS | POS |
| Morphine-3-beta-glucuronide | POS | POS | POS |
| Morphine-6-beta-glucuronide | POS | POS | POS |
| Nadolol | NEG | POS | POS |
| Nalmefene | POS | POS | POS |
| Nalorphine | POS | POS | POS |
| Naloxone-3-beta-D-glucuronide | POS | POS | POS |
| Naltrexone | POS | POS | POS |
| Naltriben | POS | POS | POS |
| Naproxen | NEG | POS | POS |
| N-Desmethylclomipramine | NEG | POS | POS |
| N-Desmethylflunitrazepam | NEG | POS | POS |
| N-Desmethyltramadol | NEG | POS | POS |
| N-Desmethyltrimipramine | NEG | POS | POS |
| Neomycin | NEG | POS | POS |
| Nitrazepam | NEG | POS | POS |
| Norbenzoylecgonine | NEG | POS | POS |
| Norbuprenorphine | NEG | POS | POS |
| Norcocaethylene | NEG | POS | POS |
| Norcocaine | NEG | POS | POS |
| Norcodeine | POS | POS | POS |
| Nordiazepam | NEG | POS | POS |
| Norfentanyl | NEG | POS | POS |
| Nor-LAAM | NEG | POS | POS |
| Nor-LSD/Nor-ISO-LSD | NEG | POS | POS |
Table 5d: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Sys
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| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
|---|---|---|---|
| Normeperidine | NEG | POS | POS |
| Normeperidinic acid | NEG | POS | POS |
| Normorphine | POS | POS | POS |
| Noroxycodone | POS | POS | POS |
| Noroxymorphone | POS | POS | POS |
| Norpropoxyphene | NEG | POS | POS |
| Nortriptyline | NEG | POS | POS |
| Noscapine | NEG | POS | POS |
| O-Desmethyltramadol | NEG | POS | POS |
| Oxazepam | NEG | POS | POS |
| Oxprenolol | NEG | POS | POS |
| Oxycodone | POS | POS | POS |
| Oxymorphone | POS | POS | POS |
| p-Acetamidophenyl-beta-D-glucuronide | NEG | POS | POS |
| Papaverine | NEG | POS | POS |
| Pemoline | NEG | POS | POS |
| Penicillin G | NEG | POS | POS |
| Pentazocine | NEG | POS | POS |
| Pentobarbital | NEG | POS | POS |
| Perphenazine | NEG | POS | POS |
| Phendimetrazine | NEG | POS | POS |
| Phenelzine | NEG | POS | POS |
| Phenobarbital | NEG | POS | POS |
| Phenothiazine | NEG | POS | POS |
| Phentermine | NEG | POS | POS |
| Phenylbutazone | NEG | POS | POS |
| Phenylethyamine | NEG | POS | POS |
| Phenylpropanolamine | NEG | POS | POS |
| Phencyclidine | NEG | POS | POS |
| R,R (-)-Pseudoephedrine | NEG | POS | POS |
| Phencyclidine Morpholine | NEG | POS | POS |
| PMA | NEG | POS | POS |
| PMMA | NEG | POS | POS |
| Prednisolone | NEG | POS | POS |
Table 5d: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Sys
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| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
|---|---|---|---|
| Prilocaine | NEG | POS | POS |
| Prochlorperazine | NEG | POS | POS |
| Progesterone | NEG | POS | POS |
| Promazine | NEG | POS | POS |
| Promethazine | NEG | POS | POS |
| Propiomazine | NEG | POS | POS |
| Propionylpromazine | NEG | POS | POS |
| Propoxyphene | NEG | POS | POS |
| Protriptyline | NEG | POS | POS |
| Quinidine | NEG | POS | POS |
| R (+) Methcathinone | NEG | POS | POS |
| R (-) Epinephrine | NEG | POS | POS |
| R (+) Cathinone | NEG | POS | POS |
| Salbutamol | NEG | POS | POS |
| Secobarbital | NEG | POS | POS |
| Sertraline | NEG | POS | POS |
| Stanazalol | NEG | POS | POS |
| Streptomycin | NEG | POS | POS |
| Sulfadimethoxine | NEG | POS | POS |
| Sulfamethazine | NEG | POS | POS |
| Sulfathiazole | NEG | POS | POS |
| Temazepam | NEG | POS | POS |
| Terbutaline | NEG | POS | POS |
| Tetracycline | NEG | POS | POS |
| Thebaine | POS | POS | POS |
| Theophylline | NEG | POS | POS |
| Thioridazine | NEG | POS | POS |
| Tramadol | NEG | POS | POS |
| Triamcinolone | NEG | POS | POS |
| Triazolam | NEG | POS | POS |
| Trifluoperazine | NEG | POS | POS |
| Trifluopromazine | NEG | POS | POS |
| Trimeprazine | NEG | POS | POS |
| Trimipramine | NEG | POS | POS |
Table 5d: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Sys
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| Compound | -50% CO(150pg/mg) | +125% CO(375pg/mg) | +150% CO(450pg/mg) |
|---|---|---|---|
| Tylosin | NEG | POS | POS |
| Tyramine | NEG | POS | POS |
| Yohimbic acid | NEG | POS | POS |
| Yohimbine | NEG | POS | POS |
| Zolpidem | NEG | POS | POS |
| Zopiclone | NEG | POS | POS |
Table 5d: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Sys
This study demonstrated that the presence of the structurally similar compounds Buprenorphine, Noroxymorphone, 3-Methoxynaltrexone, Morphine-6-beta-glucuronide, Nalmefene, Nalorphine, Naloxone-3-beta-D-glucuronide, Naltriben, and Noroxymorphone, may contribute to an Opiate or Oxycodone positive ELISA result when utilizing this protocol.
To better under the observed cross-reactivity of the above identified compounds, the concentration ranges were extended to generate additional cross-reactivity (CR) data. With the extremely high concentrations, some cross-reactivity was now observable in all except for Nalmefene, Naloxone-3-beta-D-glucuronide and Naltriben.
The interference tests were also rerun at -50% of cutoffs. There was good correlation with the previous dataset. One interference test produced a different outcome. Specifically, Noroxymorphone in opiates assay. The calculated equivalents based on the CR curve resulted in a calculated equivalent concentration close to the assay cutoff.
| Compound | Concentration ofCompound (pg/mg)Equivalent to 300 pg/mgOpiates Cutoff Control | Percent Cross-Reactivity (%) |
|---|---|---|
| Morphine-6-β-D-glucuronide | 800 | 37.5 |
| Nalorphine | 8500 | 3.5 |
| Buprenorphine | 600000 | 0.050 |
| 3-Methoxynaltrexone | 900000 | 0.033 |
| Naltrexone | 3000000 | 0.010 |
| Noroxymorphone | 4000000 | 0.008 |
| Nalmefene | * | 0.00 |
| Naloxone-3-β-D-glucuronide | * | 0.00 |
| Naltriben | * | 0.00 |
5e: Cross reactivity of Opiates ELISA with Structurally Similar Compounds
- Unable to generate an assay absorbance equivalent to 300pg/mg Opiates cutoff. Highest concentration tested was 4,000,000pg/mg.
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| Compounds | ELISA Assay Results | Opiates equivalents (pg/mg) |
|---|---|---|
| Morphine 6-β-D-glucuronide | POS | 150300 |
| Nalorphine | POS | 14300 |
| Buprenorphine | POS | 500 |
| 3-Methoxynaltrexone | NEG | 432 |
| Naltrexone | NEG | 340 |
| Noroxymorphone | NEG | 332 |
| Nalmefene | NEG | 332 |
| Naloxone 3-β-D-glucuronide | NEG | 332 |
| Naltriben | NEG | 332 |
Table 5f: Interferences of Structurally Related and Unrelated Compounds on Opiates ELISA Assay All results at the -50% of cutoff
Table 5g: Cross Reactivity of Omega Laboratories, Inc. Oxycodone ELISA with Structurally Similar Compounds
| Compound | Concentration of Compound(pg/mg) Equivalent to 300pg/mg Oxycodone CutoffControl | Percent Cross-Reactivity (%) |
|---|---|---|
| Morphine-6-β-D-glucuronide | 31000 | 0.97 |
| Nalorphine | 300000 | 0.10 |
| Buprenorphine | * | 0.00 |
| 3-Methoxynaltrexone | 380000 | 0.08 |
| Naltrexone | 50000 | 0.60 |
| Noroxymorphone | 58000 | 0.52 |
| Nalmefene | 300000 | 0.10 |
| Naloxone-3-β-D-glucuronide | 680000 | 0.04 |
| Naltriben | 300000 | 0.10 |
- Unable to generate an assay absorbance equivalent to 300pg/mg Oxycodone cutoff. Highest concentration tested was 4,000,000pg/mg
Table 5h: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Assay All results at the -50% of cutoff
| Compound | ELISA AssayResults | Oxycodone equivalents(pg/mg) |
|---|---|---|
| Morphine 6-β-D-glucuronide | POS | 4,180 |
| Nalorphine | POS | 700 |
| Buprenorphine | NEG | 340 |
| 3-Methoxynaltrexone | POS | 620 |
{31}------------------------------------------------
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| Compound | ELISA AssayResults | Oxycodone equivalents(pg/mg) |
|---|---|---|
| Naltrexone | POS | 2,700 |
| Noroxymorphone | POS | 2,380 |
| Nalmefene | POS | 700 |
| Naloxone 3-β-D-glucuronide | POS | 460 |
| Naltriben | POS | 340 |
Table 5h: Interferences of Structurally Related and Unrelated Compounds on Oxycodone ELISA Assay All results at the -50% of cutoff
None of the other compounds studied demonstrated any interference with the protocol.
CALIBRATOR AND CONTROL:
The Omega Laboratories, Inc. ELISA Opiates and Oxycodone Screening Protocols utilize in-house prepared calibrators and control solutions. The study successfully demonstrated the validation and stability of these solutions and the traceability to NIST standards.
The data demonstrating the stability of morphine and oxycodone in methanol for a period of one year when stored refrigerated in an amber bottle was provide as part of K103161. The quantitative values of 271 and 291 pg/mg for morphine and oxycodone, respectively, after a one year period is within 10% of the target value of 300 pg/mg. The study validated the one 1 year expiration date for the Calibrator Stock Solution.
STABILITY:
Hair samples were taken from the head were packaged (stored) in the Omega Collection Kit (The Hair Collection Kit consists of a poly transport bag, a small piece of foil, a small specimen pouch (envelope). The Collection Kit, containing the hair sample was previously confirmed positive then stored for an average of 3.1 years for opiate samples and stored for approximately 2 years for oxycodone and hydrocodone samples.
Fifty-four samples varying in ethnic origin, hair color and curvature were tested.
| Study Observation | Morphine | Codeine | 6-AM | Oxycodone | Hydrocodone |
|---|---|---|---|---|---|
| Range in concentration pg/mghair (Before) | 520 - 1690 | 480 - 1150 | 600 - 2140 | 157 - 5174 | 196 - 2524 |
| Range in concentration pg/mghair (After) | 530 - 1588 | 539 - 1132 | 636 - 1987 | 156 - 4638 | 207 - 2359 |
| Mean Change | 1% | 7% | 4% | -4% | -5% |
| % Max and Min Decrease | --20% and -3% | --2% | -8% and -7% | -16% and -1% | -23% and -1% |
Table 6:Storage Stability Study Data Summary Ranges
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| Study Observation | Morphine | Codeine | 6-AM | Oxycodone | Hydrocodone |
|---|---|---|---|---|---|
| % Max and Min Increase | 35% and 1% | 18% and 1% | 16% and 1% | 15% and 3% | 6% and 5% |
| Number that increased inconcentration | 5 | 8 | 5 | 4 | 3 |
| Number that decreased inconcentration | 7 | 2 | 2 | 8 | 10 |
Table 6:Storage Stability Study Data Summary Ranges
Based on the data presented, opiates are stable in hair for 3 years and oxycodone and hydrocodone are stable for 2 year.
SHIPPING:
260 head hair samples were used in the shipping study; 155 samples previously confirmed positive, 100 previously screened negative samples and 5 samples that were confirmed below the 300 pg/mg cutoff. Each box contained a variety of hair color and curvature.
The minimum and maximum shipping temperature and humidity ranges are shown in the tables for the Negative samples and for the Positive samples below. Negative samples were shipped separate from the Positive samples.
| DataLogger ID | Shipped to Location ThenReturned to OmegaLaboratories | Min Temp(°C) | Max Temp (ºC) | Min Humidity(%RH) | Max Humidity(%RH) |
|---|---|---|---|---|---|
| 7310005629 | 1. Portland, Maine | -12.7 | 44.4 | 10.8 | 100 |
| 7310005644 | 2. Anchorage, Alaska | -9.4 | 44.9 | 8.1 | 96.1 |
| 7310005628 | 3. Naples, Florida | -10.8 | 43.3 | 4.4 | 100 |
| 7310005627 | 4. Tempe Arizona | -12.2 | 42.9 | 8.9 | 73.5 |
Table 7a: Negative Samples Shipping Temperatures and Humidity Ranges Negative Samples
| Table 7b: Positive Samples Shipping Temperatures and Humidity Ranges Positive Samples | |
|---|---|
| DataLogger ID | Shipped to Location ThenReturned to OmegaLaboratories | Min Temp(°C) | Max Temp (°C) | Min Humidity(%RH) | Max Humidity(%RH) |
|---|---|---|---|---|---|
| 7310005629 | 1. Portland, Maine | -12.8 | 50.8 | 0 | 97.8 |
| 7310005644 | 2. Anchorage, Alaska | -13.8 | 47.6 | 0 | 100 |
| 7310005628 | 3. Naples, Florida | -11.6 | 51.3 | 0 | 100 |
| 7310005627 | 4. Tempe Arizona | -15.3 | 41.9 | 3.2 | 100 |
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The Shipping Study demonstrated that there is no adverse effect on hair samples that would affect the screening assay when samples are exposed to extreme temperatures and variations in humidity that might occur during sample transport. The average mean % of change in screening result prior to shipping and after shipping was 1.9% for all locations combined. Four samples out of 260 had screening results that were different prior to and after shipping. All of these samples were within ±50% (150-450 pg/mg) of the cutoff level where variances in a qualitative screening assay are to be expected.
COSMETIC TREATMENT:
Numerous studies have demonstrated that the use of cosmetic treatments can reduce the amount of drugs and metabolites detected in hair specimens. This effect is completely dependent upon the nature of the hair specimen and the treatment used, and is independent of the method of analysis. This study demonstrates that the Omega Laboratories, Inc. ELISA Opiates Screening Protocol is not an exception to this phenomenon.
Test conditions:
- . BLEACH #1 - Salon Care Blue Flash Professional Powder Lightener BLEACH #2 - Loreal Super Oreal Blanc® Professional Powder Bleach
- . PERM #1 - Naturelle Natural Curls Alkiline Perm PERM #2 - Natural Apple Self-Timing Perm
- . DYE #1 - Revlon® Colorsilk™ Black DYE #2 - Garnier Herbashine Soft Mahogany Dark Brown
- . RELAXER #1 - Silk Elements™ No-Lye Sensitive Scalp Relaxer System RELAXER #2 - Ultra Precise No-Lye Conditioning Relaxer
- . SHAMPOO #1 - After Burner drug removing shampoo SHAMPOO #2 - Ultra Cleanse drug removing shampoo
176 hair samples were used in this study. Of the 176 hair specimens were identified as positive in the untreated Run No. 1 for opiates and/or oxycodone by ELISA assays and 64 specimens were identified as negative by ELISA assay. The ethnic origin, hair color and curvature were documented.
Each specimen was divided into 2 aliquots. One aliquot was analyzed by the ELISA protocol and the GC/MS confirmation method
The second aliquots were randomly assigned to the hair treatments listed above and the treatments were performed following the product insert.
Treated aliquots were analyzed by the ELISA protocol as summarized and the GC/MS confirmation method.
| Treatment | Opiates | Oxycodone/Hydrocodone | Comment |
|---|---|---|---|
| Bleach | None | None | NA |
| Dye | Pos⇒Neg (57)Neg⇒Pos (110) | None | Change at cutoff |
| Permanent | Neg⇒Pos (29)* | None | Change due to crossreactivity |
Table 8: Changes in ELISA Assay Test Results after Cosmetic Treatment (Pos ⇒ Nea or Nea ⇒ Pos)
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| Treatment | Opiates | Oxycodone/Hydrocodone | Comment |
|---|---|---|---|
| Relaxer | None | None | NA |
| Shampoo | None | Pos→Neg (89)Pos→Neg (65) | Change at cutoff |
Table 8: Changes in ELISA Assay Test Results after Cosmetic Treatment (Pos ⇒ Neg or Neg ⇒ Pos)
*Cross-reactive at 0.24 for HDC to Opiates
There was no single treatment that had a more defined effect on the assay. The single largest change was observed in the relaxer for oxycodone at -34.8%. Bleach treatment appeared to have the consistent result across all drugs. All of the treatment appeared to have the greatest change on oxycodone. See Table 9 for summary of treatment percent change review.
| Treatment | Opiates Mean change inconcentration (pg/mg) | OXY Mean change inconcentration (pg/mg) | HCD Mean change inconcentration(pg/mg) |
|---|---|---|---|
| Bleach | -10.4% | -15% | -10% |
| Permanent | -7.5 | -14.5 | -13.6 |
| Dyeing | -7.5% | -18.3% | -15.6% |
| Relaxer | -2% | -34.8% | -7% |
| Shampoo | -1.4% | -17.8% | -3.4% |
Table 9: Summary GC/MS data for Cosmetic Studies
ENVIRONMENTAL CONTAMINATION:
Two studies were performed to investigate whether confirmatory testing procedures are able to distinguish between true analytically positive samples and those that have been externally exposed to Opiates, Oxycodone and Hydrocodone. The focus of the studies was to demonstrate that a methanol wash procedure mitigates the risk of false positive results while maintaining true analytical positive results.
The first study involved exposing drug-free hair to Opiates, Oxycodone and Hydrocodone, washing the hair with methanol three times, performing confirmation testing on the samples and the washes, and observing the final test result. The second study involved performing confirmation testing on known positive samples and observing whether the methanol washes change the final result. Head hair was used for this study.
Evaluating potential environmental contamination and the effectiveness of a methanol wash using this study design, all analytically negative samples tested remained negative after being subjected to Opiates, Oxycodone and Hydrocodone by the exposure modes described followed by a single methanol wash.
Additionally, all clinically positive samples tested remain positive after the wash steps were performed.
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SUMMARY CONCLUSION:
The comparison of results of the proposed assay with the confirmatory GC/MS testing of head and body hair samples showed the results to be substantially equivalent
The candidate Omega Hair Drug Screening Assay for Opiates, Oxycodone and Hydrocodone (head and body hair) is substantially equivalent to the predicate Omega Hair Drug Screening Assay for Opiates, Oxycodone and Hydrocodone (K103161 for head hair) based on the design and performance studies discussed in this summary. Supporting Performance Testing presented for review in this document, includes agreement, precision, specificity, interference (including cosmetic effects), removal of environmental contamination, stability and shipping tests.
§ 862.3650 Opiate test system.
(a)
Identification. An opiate test system is a device intended to measure any of the addictive narcotic pain-relieving opiate drugs in blood, serum, urine, gastric contents, and saliva. An opiate is any natural or synthetic drug that has morphine-like pharmocological actions. The opiates include drugs such as morphine, morphine glucoronide, heroin, codeine, nalorphine, and meperedine. Measurements obtained by this device are used in the diagnosis and treatment of opiate use or overdose and in monitoring the levels of opiate administration to ensure appropriate therapy.(b)
Classification. Class II (special controls). An opiate test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (e.g., programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).